Associations of Metabolic Syndrome with Total Testosterone and Homocysteine Levels in Male Korean Workers

BACKGROUND: Low testosterone is associated with metabolic syndrome (MetS), and homocysteine (Hcy) is elevated in individuals with MetS. We investigated the relationships of total testosterone (TT) and serum Hcy levels with MetS in male Korean workers. METHODS: We conducted a cross-sectional study including 8,606 male workers, aged 20 to 58 years, who underwent a physical examination in 2015. MetS was diagnosed based on the criteria of the 2009 harmonized definition, while the Korean standard for waist circumference (WC) was used. Participants' biochemical parameters, including TT and serum Hcy, were measured, and participants were divided into quartiles. Multiple logistic regression models were used to estimate the association of MetS and its individual components depending on TT and serum Hcy quartiles. RESULTS: The prevalence of MetS in the study population was 16%. TT was lower in participants with MetS than in those without MetS (P<0.001). By contrast, Hcy level was similar between groups (P=0.694). In multiple logistic regression analysis, the odds ratio for the lowest TT quartile was 1.29 (95% confidence interval, 1.06 to 1.57) after adjusting for potential confounders. Participants with lower TT were more likely to have high WC, hypertriglyceridemia, and low high density lipoprotein levels. Serum Hcy levels were not significantly associated with MetS. Of the five components of MetS, only WC was significantly associated with serum Hcy. CONCLUSION: In male Korean workers, TT may be an independent predictor of MetS, and serum Hcy levels could be a marker of abdominal obesity. However, future prospective studies are needed.

Loss of Mismatched HLA Detected in the Peripheral Blood of an AML Patient who Relapsed After Haploidentical Hematopoietic Stem Cell Transplantation

No abstract available.

Changes in 18F-Fluorodeoxyglucose Uptake in the Spinal Cord in a Healthy Population on Serial Positron Emission Tomography/Computed Tomography / 전남의대학술지

We aimed to determine the changes in 18F-fluorodeoxyglucose (FDG) uptake in the spinal cord on two serial positron emission tomography/computed tomography (PET/CT) scans in a healthy population. We retrospectively enrolled healthy people who underwent PET/CT twice for cancer screening. We excluded those who had degenerative vertebral disease, neurologic disease, or a history of a vertebral operation. The standardized uptake value (SUVmax) of the spinal cord of each mid-vertebral body was obtained by drawing a region of interest on an axial image of PET/CT. For analysis, the cord-to-background ratio (CTB) was used (CTB=SUVmax of each level/SUVmax of L5 level). Differences in pattern, sex, age, and intervals of the two serial PET/CT scans were analyzed. A total of 60 PET/CT images of 30 people were analyzed. The mean interval between the two PET/CT imaging studies was 2.80+/-0.94 years. On the follow-up PET/CT, significant change was shown only at the level of the C6 and T10 vertebrae (p<0.005). Mean CTB showed a decreasing pattern from cervical to lumbar vertebrae. There were two peaks at the lower cervical level (C4-6) and at the lower thoracic level (T12). Neither sex nor age significantly affected CTB. The FDG uptake of the spinal cord changed significantly on follow-up PET/CT only at the level of the C6 and T10 vertebrae. This finding is valuable as a baseline reference in the follow-up of metabolic changes in the spinal cord.

Changes in 18F-Fluorodeoxyglucose Uptake in the Spinal Cord in a Healthy Population on Serial Positron Emission Tomography/Computed Tomography / 전남의대학술지

We aimed to determine the changes in 18F-fluorodeoxyglucose (FDG) uptake in the spinal cord on two serial positron emission tomography/computed tomography (PET/CT) scans in a healthy population. We retrospectively enrolled healthy people who underwent PET/CT twice for cancer screening. We excluded those who had degenerative vertebral disease, neurologic disease, or a history of a vertebral operation. The standardized uptake value (SUVmax) of the spinal cord of each mid-vertebral body was obtained by drawing a region of interest on an axial image of PET/CT. For analysis, the cord-to-background ratio (CTB) was used (CTB=SUVmax of each level/SUVmax of L5 level). Differences in pattern, sex, age, and intervals of the two serial PET/CT scans were analyzed. A total of 60 PET/CT images of 30 people were analyzed. The mean interval between the two PET/CT imaging studies was 2.80+/-0.94 years. On the follow-up PET/CT, significant change was shown only at the level of the C6 and T10 vertebrae (p<0.005). Mean CTB showed a decreasing pattern from cervical to lumbar vertebrae. There were two peaks at the lower cervical level (C4-6) and at the lower thoracic level (T12). Neither sex nor age significantly affected CTB. The FDG uptake of the spinal cord changed significantly on follow-up PET/CT only at the level of the C6 and T10 vertebrae. This finding is valuable as a baseline reference in the follow-up of metabolic changes in the spinal cord.

Performance evaluation of the HepB Typer-Entecavir kit for detection of entecavir resistance mutations in chronic hepatitis B

BACKGROUND/AIMS: Molecular diagnostic methods have enabled the rapid diagnosis of drug-resistant mutations in hepatitis B virus (HBV) and have reduced both unnecessary therapeutic interventions and medical costs. In this study we evaluated the analytical and clinical performances of the HepB Typer-Entecavir kit (GeneMatrix, Korea) in detecting entecavir-resistance-associated mutations. METHODS: The HepB Typer-Entecavir kit was evaluated for its limit of detection, interference, cross-reactivity, and precision using HBV reference standards made by diluting high-titer viral stocks in HBV-negative human serum. The performance of the HepB Typer-Entecavir kit for detecting mutations related to entecavir resistance was compared with direct sequencing for 396 clinical samples from 108 patients. RESULTS: Using the reference standards, the detection limit of the HepB Typer-Entecavir kit was found to be as low as 500 copies/mL. No cross-reactivity was observed, and elevated levels of various interfering substances did not adversely affect its analytical performance. The precision test conducted by repetitive analysis of 2,400 replicates with reference standards at various concentrations showed 99.9% agreement (2398/2400). The overall concordance rate between the HepB Typer-Entecavir kit and direct sequencing assays in 396 clinical samples was 99.5%. CONCLUSIONS: The HepB Typer-Entecavir kit showed high reliability and precision, and comparable sensitivity and specificity for detecting mutant virus populations in reference and clinical samples in comparison with direct sequencing. Therefore, this assay would be clinically useful in the diagnosis of entecavir-resistance-associated mutations in chronic hepatitis B.

De Novo Superinfection of Hepatitis B Virus in an Anti-HBs Positive Patient with Recurrent Hepatitis C Following Liver Transplantation

A 60-year-old woman with end stage liver cirrhosis caused by genotype 2 hepatitis C virus (HCV) infection received an orthotopic liver transplantation (OLT). The patient was negative for the hepatitis B surface antigen (HBsAg) and positive for the anti-hepatitis B surface antibody (anti-HBs) prior to and one and a half months following the OLT. Due to reactivation of hepatitis C, treatment with interferon-alpha and Ribavirin started two months following the OLT and resulted in a sustained virological response. We performed a liver biopsy because a biochemical response was not achieved. Surprisingly, liver pathology showed HBsAg-positive hepatocytes with a lobular hepatitis feature, which had been negative in the liver biopsy specimen obtained one and a half months post-OLT. High titers of both HBsAg and HBeAg were detected, while anti-HBs antibodies were not found. Tests for IgM anti-hepatitis B core antibody and anti-delta virus antibodies were negative. The serum HBV DNA titer was over 1x10(7) copies/mL. A sequencing analysis showed no mutation in the "a" determinant region, but revealed a mixture of wild and mutant strains at an overlapping region of the S and P genes (S codon 213 (Leu/Ile); P codons 221 (Phe/Tyr) and 222 (Ala/Thr)). These findings suggest that de novo hepatitis B can develop in patients with HCV infection during the post-OLT period despite the presence of protective anti-HBs.

Effect on complications associated with its position and angulation following mandibular third molar extraction

INTRODUCTION: Mandibular third molar extraction is one of the most common procedures performed in oral and maxillofacial surgery units. Although the overall complication rate is low with most complications being minor, mandibular third molar removal is so common that the population morbidity of complications might be significant. Therefore, efforts to limit intraoperative or postoperative complications might have a significant impact in terms of enhancing the patient outcome. The aims of this study were to identify the position and angulation associated complications after mandibular third molar extractions. MATERIALS AND METHODS: This study surveyed 568 patients who had a mandibular third molar extracted, showed clinical complications and underwent a radiographic measurement of the available space, depth and spatial relationship. RESULTS: The results obtained were as follows: 1. The complications were a dry socket, nerve injury, root rest, infection, bleeding, hamatoma, and adjacent teeth injury. 2. There were no significant differences between the complication and ramus relationship (available space) of the mandibular third molar. 3. There were no significant differences between the complications and depth of the mandibular third molar. 4. There were no significant differences between the complications and spatial relationship of the mandibular third molar. CONCLUSION: There were no significant differences in the complication rate, ramus relationship, depth and spatial relationship of the mandibular third molar. This suggests that the position and angulation of the mandibular third molar may not have an impact on the complications. The relationship between the position and angulation of the mandibular third molar, and complications deserves a further study using longitudinal data.

Association of Methylenetetrahydrofolate Reductase (MTHFR 677C>T and 1298A>C) Polymorphisms and Haplotypes with Silent Brain Infarction and Homocysteine Levels in a Korean Population

PURPOSE: Methylenetetrahydrofolate reductase (MTHFR) is the main regulatory enzyme for homocysteine metabolism. In the present study, we evaluated whether the MTHFR 677C>T and 1298A>C gene polymorphisms are associated with SBI and plasma homocysteine concentration in a Korean population. MATERIALS AND METHODS: We enrolled 264 patients with SBI and 234 healthy controls in South Korea. Fasting plasma total homocysteine (tHcy) concentrations were measured, and genotype analysis of the MTHFR gene was carried out. RESULTS: The plasma tHcy levels were significantly higher in patients with SBI than in healthy controls. Despite a significant association between the MTHFR 677TT genotype and hyperhomocysteinemia, the MTHFR 677C>T genotypes did not appear to influence susceptibility to SBI. However, odds ratios of the 1298AC and 1298AC + CC genotypes for the 1298AA genotype were significantly different between SBI patients and normal controls. The frequencies of 677C-1298A and 677C-1298C haplotypes were significantly higher in the SBI group than in the control group. CONCLUSION: This study demonstrates that the MTHFR 1298A>C polymorphism is a risk factor for SBI in a Korean population. The genotypes of 677C>T and 1298A>C polymorphisms interact additively, and increase the risk of SBI in Korean subjects.

A YIDD Mutation in a Case of Recurrent Hepatitis B after Liver Transplantation Induced by an S-escape Mutant

A 47-year-old woman underwent orthotopic liver transplantation (OLT) for hepatitis B virus (HBV)-related end-stage liver cirrhosis. The patient received hepatitis B immunoglobulin prophylaxis after OLT. Despite the protective level of the serum anti-hepatitis-B surface antibody, HBV recurred at 22 months post-OLT and induced subacute hepatic failure. The pre-OLT HBV genome contained a complex mutation pattern in overlapping frame regions of the surface (S) and polymerase (P) genes, which is the same mutation pattern as seen in post-OLT HBV DNA. G145R and K141R mutations in the "a" determinant were detected only in the post-OLT sample. Clevudine (30 mg once daily) was administered for recurrent hepatitis B. Hepatitis B was reactivated with a flare-up, and a M204I mutation (YIDD mutant type) appeared with a higher viral load at 9 months after clevudine treatment. We report here a case of a YIDD mutation that developed in recurrent hepatitis B after OLT induced by an S-escape mutant.

Virologic Response at 12 Months of Treatment Predicts Sustained Antiviral Efficacy in Patients with Adefovir-Treated Lamivudine-Resistant Chronic Hepatitis B

BACKGROUND/AIMS: The aim of our study was to define the potential role of virologic response at 12 months of treatment (VR12) in predicting subsequent virologic and clinical outcomes in adefovir (ADV)-treated lamivudine-resistant chronic hepatitis B. METHODS: Two hundred and four patients with lamivudine-resistant chronic hepatitis B virus (HBV) treated with ADV monotherapy were included. Serum HBV DNA was quantified by real-time polymerase chain reactions. VR12 was defined as a HBV DNA level of less than 4 log10 copies/mL after 12 months of ADV treatment. RESULTS: VR12 was observed in 110 of the 204 patients (54%). The mean HBV DNA reductions from baseline after 12 months of ADV treatment were 3.8 and 1.9 log10 copies/mL in patients with and without VR12, respectively (p<0.001). The hepatitis B "e" antigen (HBeAg) seroconversion rates in patients with and without VR12 were 32% and 14% at 12 months treatment, respectively (p=0.018), and 40% and 27% at 24 months of treatment (p=0.032). The genotypic mutation rates to ADV in patients with and without VR12 were 0% and 6% at 12 months of treatment, respectively (p=0.033), and 21% and 42% at 24 months (p=0.012). The rates of viral breakthrough in patients with and without VR12 were 0% and 7% at 12 months of treatment, respectively (p=0.072), and 9% and 25% at 24 months (p=0.006). CONCLUSIONS: Patients without VR12 may need to switch to or add on other potent antiviral drugs in their medical regimens.

Genotypic resistance to entecavir in chronic hepatitis B patients / 대한간학회지

BACKGROUND/AIMS: The prevalence and clinical characteristics of entecavir (ETV) resistance is not well known. The aim of this study was to determine the frequency of genotypic resistance in nonresponders and virologic breakthrough (VBT) patients. METHODS: The medical records of 76 chronic hepatitis B patients treated for a least 6 months from October 2006 to October 2008 were reviewed retrospectively. We divided patients into two groups: nucleoside analogue (NA)-naive patients (n=38) and LAM experienced patients (n=38). NA-naive and LAM experienced patients received ETV at 0.5 and 1.0 mg/day, respectively. The virologic response and VBT were investigated in both groups. We used the multiplex restriction fragment mass polymorphism (RFMP) method to test genotypic resistance at the rtI169, rtT184, rtS202, rtM204, and rtM250 sites. RESULTS: Age, gender, serum ALT, and HBV DNA level before treatment did not differ between the groups. Neither VBT nor nonresponse was observed in the NA-naive group, whereas VBT and nonresponse were observed in three patients each in the lamivudine (LAM)-experienced group; all six patients had YMDD mutation at study enrollment, all three patients with VBT had genotypic resistance to ETV, but the three nonresponse patients did not have genotypic resistance to ETV. CONCLUSIONS: We suspect that VBT is mostly associated with genotypic resistance to ETV. However, nonresponse might be associated with the continuance or reselection of the YMDD mutant in LAM-experienced patients.

18F-FDG PET/CT Finding in a Case of Xanthogranulomatous Pyelonephritis / 핵의학분자영상

Xanthogranulomatous pyelonephritis is an uncommon chronic renal infection, which is usually found on middle-aged women and is rare in infant. Sometimes it forms focal mass like lesion of kidney with pathologically characteristic lipid-laden macrophage. A 1-month female infant was admitted for fever and moaning sound. On work-up of urinary tract infection, abdomen ultrasonography and computed tomography revealed a large mass on the upper portion of right kidney and PET/CT showed homogeneously increased 18F-FDG uptake. The radical nephrectomy of right kidney was performed and histology revealed a focal xanthogranulomatous pyelonephritis. To our knowledge, this is the first report presenting the finding of 18F-FDG PET/CT in the childhood xanthogranulomatous pyelonephritis.

Ipsilateral Cerebral and Contralateral Cerebellar Hyperperfusion in Patients with Unilateral Cerebral Infarction; SPM Analysis / 핵의학분자영상

PURPOSE: Cortical reorganization has an important role in the recovery of stroke. We analyzed the compensatory cerebral and cerebellar perfusion change in patients with unilateral cerebral infarction using statistical parametric mapping (SPM). MATERIALS AND METHODS: Fifty seven (99m)Tc-Ethylene Cystein Diethylester (ECD) cerebral perfusion SPECT images of 57 patients (male/female=38/19, mean age=56+/-17 years) with unilateral cerebral infarction were evaluated retrospectively. Patients were divided into subgroups according to the location (left, right) and the onset (acute, chronic) of infarction. Each subgroup was compared with normal controls (male/female=11/1, mean age=36+/-10 years) in a voxel-by-voxel manner (two sample t-test, p<0.001) using SPM. RESULTS: All 4 subgroups showed hyperperfusion in the ipsilateral cerebral cortex, but not in the contralateral cerebral cortex. Chronic left and right infarction groups revealed hyperperfusion in the ipsilateral primary sensorimotor cortex, meanwhile, acute subgroups did not. Contralateral cerebellar hyperperfusion was also demonstrated in the chronic left infarction group. CONCLUSION: Using (99m)Tc-ECD SPECT, we observed ipsilateral cerebral and contralateral cerebeller hyperperfusion in patients with cerebral infarction. However, whether these findings are related to the recovery of cerebral functions should be further evaluated.

Clinical Effectiveness of Restriction Fragment Mass Polymorphism Assay for Human Papillomavirus Genotyping / 임상검사와정도관리

BACKGROUND: Human papillomavirus (HPV) infection is the main cause of cervical cancer and with the advent of genotype specific vaccines, there is increased need for accurate, broad-spectrum and high-throughput methods for HPV genotyping. A MALDI-TOF mass spectrometry (MS)-based restriction fragment mass polymorphism (RFMP) assay has proven to accurately and reliably genotype a wide variety of HPV. METHODS: We evaluated the clinical utility of the RFMP assay in HPV genotyping by testing a total of 2,689 specimens taken from liquid-based cytology, which was composed of normal cytology, atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL) and invasive squamous cervical cancer (SCC). RESULTS: Overall HPV positive rate of total specimens was 32.5% and the high-risk positivity was 16.4%. The HPV positive rates were increased as increasing severity level of cervical lesion. Predominant high-risk HPV genotypes were found as following order; 52 (18.6%), 16 (13.7%), 18 (3.8%), 58 (3.4%), 56 (2.6%) and 31 (2.5%). The high-risk HPV positivities according to cytologic diagnosis were 10.7% (238/2229), 31.7% (76/240), 50.0% (88/176), 86.0% (37/43), 100% (1/1) in normal, ASCUS, LSIL, HSIL and SCC subgroups, respectively. The concordance rate and Kappa value between sequencing and RFMP assays were 96.6% and 0.932 (95%CI: 0.908-0.956). CONCLUSIONS: The RFMP HPV genotyping assays showed high concordance with sequencing. The assay is simple, and can accurately detect and identify HPV genotypes in samples with various levels of cytological lesions. The results demonstrated that RFMP assay should be clinically suitable for HPV genotyping in laboratories.

Factors affecting initial virologic response and emergence of resistant mutants after adefovir treatment in lamivudine-resistant chronic hepatitis B patients / 대한간학회지

BACKGROUND/AIMS: Adefovir dipivoxil (adefovir) effectively inhibits both wild-type and lamivudine-resistant hepatitis B virus (HBV) replication. The development of adefovir resistance is both delayed and infrequent compared with lamivudine resistance. The aim of this study was to characterize the serologic, biochemical, and virologic response to adefovir, and to explore the factors affecting initial virologic response (IVR, defined as a decrease in serum HBV below 4 log10copies/mL after 6 month of treatment) and adefovir resistance in lamivudine resistant HBV-infected patients. METHODS: This study population comprised 76 patients with lamivudine-resistance who had received adefovir for more than 12 months between March 2004 and December 2006. The adefovir-resistant mutant was assayed at 6 months and 12 months during adefovir administration. Restriction-fragment mass polymorphism analysis was used for detecting YMDD and adefovir mutants. RESULTS: After adefovir administration, an IVR was observed in 31% of the patients with lamivudine resistance. Factors associated with an IVR were HBeAg negativity (P=0.04) and the presence of liver cirrhosis (P=0.04). Age, sex, pretreatment levels of alanine aminotransferase and aspartate aminotransferase, pretreatment HBV DNA levels, presence of precore mutation, and type of YMDD mutants were not related to an IVR during adefovir treatment. The prevalence of adefovir resistance was 5% and 13% at 6 months and 12 months after therapy, respectively. Mixed infection of the precore mutant was a risk factors for the emergence of adefovir resistance (P=0.01). CONCLUSIONS: Lamivudine-resistant HBV patients exhibiting HBeAg negativity and liver cirrhosis were more likely to achieve an IVR after adefovir therapy. Adefovir resistance was associated with mixed infection of the precore mutant.

Influence of Combined Methionine Synthase (MTR 2756A > G) and Methylenetetrahydrofolate Reductase (MTHFR 677C > T) Polymorphisms to Plasma Homocysteine Levels in Korean Patients with Ischemic Stroke

PURPOSE: Methionine synthase (MTR) and 5,10-methylenetetrahydrofolate reductase (MTHFR) are the main regulatory enzymes for homocysteine metabolism. The present case- control study was conducted to determine whether there is an association between the MTR 2756A > G or MTHFR 677C > T polymorphism and plasma homocysteine concentration in Korean subjects with ischemic stroke. MATERIALS AND METHODS: DNA samples of 237 patients who had an ischemic stroke and 223 age and sex-matched controls were studied. MTR 2756A > G and MTHFR 677C > T genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Frequencies of mutant alleles for MTR and MTHFR polymorphisms were not significantly different between the controls and cases. The patient group, however, had significantly higher homocysteine concentrations of the MTR 2756AA and MTHFR 677TT genotypes than the control group (p=0.04 for MTR, p=0.01 for MTHFR). The combined MTR 2756AA and MTHFR 677TT genotype (p= 0.04) and the homocysteine concentrations of the patient group were also higher than those of the controls. In addition, the genotype distribution was significant in the MTHFR 677TT genotype (p=0.008) and combined MTR 2756AA and MTHFR 677TT genotype (p=0.03), which divided the groups into the top 20% and bottom 20% based on their homocysteine levels. CONCLUSION: The results of the present study demonstrate that the MTR 2756A > G and MTHFR 677C > T polymorphisms interact with elevated total homocysteine (tHcy) levels, leading to an increased risk of ischemic stroke.

Human Papilloma Virus Genotyping Assay using Restriction Fragment Mass Polymorphism Analysis, and Its Comparison with Sequencing and Hybrid Capture Assays / 대한진단검사의학회지

BACKGROUND: Infection with human papilloma virus (HPV) is the main cause of cervical cancer, and HPV genotyping is of increasing importance for determining clinical course and management of the disease based on the HPV genotypes. Here, we established a novel matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF MS) assay, termed restriction fragment mass polymorphism (RFMP) that is suitable for genotyping multiple HPV in an accurate and high-throughput manner. We evaluated the performance of the RFMP assay in HPV genotyping by comparing the results with those of direct or clonal sequencing and hybrid capture (HC) assays. METHODS: The study population consisted of 50 patients with histologically confirmed cervical lesions and a positive test for HPV DNA. HPV genotyping was performed with RFMP, sequencing, and HC assays. The assigned genotypes and risk groups were compared among the methods. RESULTS: Concordance rates in the genotype level between RFMP vs sequencing, sequencing vs HC, and HC vs RFMP were 98% (49/50), 88% (44/50), and 88% (44/50), respectivley. Especially, RFMP and sequencing were 100% concordant when assigned high-risk group was considered identical in 1 case of mixed genotypes identified only in RFMP. The observed discrepancy between HC and the other two methods is due to the assignment of six cases of low, intermediate, or unassigned risk genotypes as high-risk group in HC method. CONCLUSIONS: RFMP, sequencing, and HC assays were highly concordant with each other in HPV genotyping. Compared to sequencing assay, RFMP assay is found to be advantageous in detecting mixed genotype infections. The accuracy and amenability to high-throughput analysis should make the RFMP assay suitable for reliable screening of HPV genotypes in clinical laboratories.

Comparison of Efficacy of Human Papilloma Virus Genotyping Assays using Restriction Fragment Mass Polymorphism and DNA Chip Analysis in Patients with Abnormal Pap Smear and Uterine Cervical Cancer

BACKGROUND: High-risk human papilloma virus (HPV) infection is the primary cause of cervical cancer; there is a need for more sensitive and reliable methods for HPV genotyping to use as screening tools for early detection and intervention. METHODS: A novel MALDI-TOF MSbased assay, termed Restriction Fragment Mass Polymorphism (RFMP) was developed for multiple HPV genotyping. Its performance was compared with DNA chip technology. The study was based on 164 cases classified as normal (n=40), ASCUS (n=53) and invasive squamous cell carcinoma (SCC, n=71) by a PAP smear and/or cervical colposcopic biopsy. RESULTS: High-risk genotypes were detected in 7.5%, 47.2% and 97.2% in normal, ASCUS and SCC groups by RFMP, and in 20.0%, 41.5% and 90.1% using DNA chip technology, respectively. The results showed substantial concordance, with a kappa coefficient of 0.688, between the methods. Diagnostic sensitivity and specificity for cervical cancer were found to be 97.2% and 92.2% with RFMP and 90.1% and 80.0% using DNA chip microarrays. CONCLUSIONS: RFMP and DNA chip technologies were shown to be reliable methods for HPV genotyping with a high concordance. The improved sensitivity and specificity should make RFMP a viable option for the management of women with cervical neoplastic lesions.

Single Nucleotide Polymorphisms in the 3'-Untranslated Region of Vascular Endothelial Growth Factor in Diabetic Retinopathy

PURPOSE: Recent studies suggest that increased expression of the vascular endothelial growth factor (VEGF) may play a role in the pathogenesis of diabetic complications. The aim of this study was to assess the potential association of VEGF gene polymorphisms with disease expression of retinopathy in patients with diabetes mellitus. METHODS: We studied 96 diabetic patients to determine whether there is an association between diabetic retinopathy in VEGF and VEGFR gene; all patients were unrelated Korean individuals (PDR, n=32; NPDR, n=25; no DMR, n=39). We analyzed VEGF and VEGFR SNP by restriction fragment mass polymorphism (RFMP). PCR was performed with primers designed to introduce a type IIS restriction endonuclease recognition sequence (FokI, BstF5I) ahead of the polymorphism site. The enzymatic cleavage of the products resulted to excision of two oligonucleotide fragments containing the variation site, and then masses of the resulting oligonucleotide fragments were examined by matrix assisted laser desorption time of flight mass spectrometry (MALDI-TOF MS). Difference are seen as the presence, absence, or mass change in peaks corresponding to the fragments affected by existence of polymorphism that have base substitution at the site of variation. RESULTS: We found that the genotype frequencies of two polymorphisms shown to be completely linked to each other (rs3025039; vf1, rs3025040; vf2) and one polymorphism (rs3812867; vfr1) within 3'-untranslated regions (3'-UTR) of the VEGF and VEGFR1 significantly differed between patients with and without retinopathy. The frequencies of the vf1, vf2 and vfr1 did not differ significantly between the NPDR and PDR group CONCLUSIONS: These data implicate the polymorphisms in the 3'-UTR of the VEGF and VEGFR1 genes as risk factors for diabetic retinopathy.

The Comparison of Restriction Fragment Mass Polymorphism with Sequencing Method for the Hepatitis C Virus Genotyping / 대한진단검사의학회지

BACKGROUND: Hepatitis C virus (HCV) genotyping is of increasing the importance in the progression to liver cirrhosis and the outcome of antiviral therapy. Restriction fragment mass polymorphism (RFMP) method which is developed recently for HCV genotyping is known to report accurate result in mixed infection. We evaluated the performance of RFMP in HCV genotyping by comparing the result of direct sequencing. METHODS: Forty-three chronically HCV infected patients in Asan Medical Center were enrolled. HCV genotyping was performed by both RFMP and direct sequencing. The results were compared from the genotype level to the subtype level. RESULTS: In the genotype level, all the results (100%) were concordant. At the subtype level, however, 2 results (2/43, 4.7%) were disconcordant. Two cases were reported as single infection of type 1b by direct sequencing while RFMP reported as mixed infection of 1b with 1a and 1b with 1c, respectively. CONCLUSIONS: Both sequencing and RFMP assays were highly concordant. We suggest that RFMP is a novel method in HCV genotyping superior to sequencing or hybridization method especially in mixed infection.